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Hydrogen sulfide reduces histopathology and improves functional outcome after traumatic brain injury in mice
Abstracts/Nitric Oxide 47 (2015) S14–S60
MnSOD. Interestingly, hypoxia induced a transient increase of MnSOD activity and mRNA expression in 30 min, followed by a final loss of MnSOD function, while the conjugate pre-treatment could timedependently improve MnSOD activity, as well as its gene expression. We also found the protective effects were mediated by STAT3 phosphorylation, for STAT3 inhibition and knockdown both attenuated the protective effects of the conjugate. Furthermore, burst of mitochondrial ROS (mROS), dispersion of mitochondrial membrane potential (ΔΨm) and fragmentation of myofibril were also ameliorated with the conjugate treatment; again, the cell-protective effects of the conjugate were canceled by the STAT3 inhibitor 5,15-DPP. Innovation and conclusion: We present a novel hydrogen sulfide donor with excellent cardio-protective potential and better watersolubility, and find the cardio-protective mechanisms. The conjugate suppresses mitochondrial ROS through activation of the STAT3– MnSOD pathway, which is a novel way of mitochondrial ROS elimination. Keywords: Leonurine–SPRC conjugate; acute myocardial infarction; mitochondrial ROS; cardiac protection; STAT3–MnSOD pathway http://dx.doi.org/10.1016/j.niox.2015.02.048
PP19 Hydrogen sulfide reduces histopathology and improves functional outcome after traumatic brain injury in mice Mingyang Zhang a,b, Xiping Chen a, Luyang Tao a a Department of Forensic Sciences, Soochow University, Suzhou, China b Department of Forensic Sciences, Nantong University, Nantong, China Introduction: Hydrogen sulfide (H2S) is a lipid-soluble, endogenously produced gaseous messenger molecule collectively known as gasotransmitter. Over the last several decades, gasotransmitters have emerged as potent cytoprotective mediators in various models of tissue and cellular injury. Objects: To investigate changes of H2S after traumatic brain injury and its possible role, a mouse traumatic brain injury (TBI) model was established. Methods: Expression of Cystathionine-β-synthase (CBS) mRNA as a H2S-producing enzyme in mouse brain was determined by reverse transcriptase–polymerase chain reaction (RT-PCR). This study examines the neuroprotective effects of Hydrogen sulfide by lesion volume, motor performance and Morris water maze test after TBI in mice. Results: From the results of RT-PCR, it was found that the expression of CBS was down-regulated in mouse brain cortex and hippocampus after brain injury. Western blot analysis revealed that CBS was present in normal mouse brain cortex and hippocampus. Hydrogen sulfide in the cortex and hippocampus exhibited dynamic changes after brain injury, in parallel with CBS mRNA and protein expression. Moreover, pretreatment with the H2S donor (NaHS) could protect the neuron against the injury induced by TBI. Noticeably, the H2S donor NaHS could reduce TBI-induced injury assessed with lesion volume. Conclusion: These data suggested that H2S may have a therapeutic potential against neuron damage and improves functional outcome. http://dx.doi.org/10.1016/j.niox.2015.02.049
PP20 Hydrogen sulfide offers neuroprotection on traumatic brain injury in mice through modulation of apoptosis- and autophagyregulating molecules Mingyang Zhang a,b, Xiping Chen a, Luyang Tao a a Department of Forensic Sciences, Soochow University, Suzhou, China b Department of Forensic Sciences, Nantong University, Nantong, China
S21
Introduction: Traumatic brain injury (TBI) is a serious public health problem affecting millions of people in the world. Hydrogen sulfide (H2S), a novel gaseous mediator, has been recognized as an important neuromodulator and neuroprotective agent in the central nervous system. Objects: To investigate the possible role of H2S in traumatic brain injury (TBI), a mouse TBI model was established. The present study was undertaken to study the effects of exogenous H2S on traumatic brain injury (TBI) and the underlying mechanisms. Methods: The effects of exogenous H2S on TBI were examined by using measurement of brain edema, behavior assessment, propidium iodide (PI) staining, and Western blotting, respectively. To investigate whether H2S can ameliorate motor and cognitive dysfunction after TBI, motor test and Morris water maze were performed. Results: Compared to TBI groups, H2S pretreatment reduced brain edema, improved motor performance and ameliorated performance in the Morris water maze test after TBI. Compared to the TBI group, H2S pretreatment could decrease the number of PI-positive cells in injured cortex, dentate gyrus, CA1 and CA3 regions. Immunoblotting results showed that H2S pretreatment reversed TBIinduced cleavage of caspase-3 and decline of Bcl-2, suppressed LC3II, Beclin-1 and Vps34 activation and maintained p62 level in injured cortex and hippocampus post TBI. Conclusions: The present study demonstrates that systemic administration of H 2 S ameliorates brain edema and behavioral symptoms in TBI models. H2S may serve as a neuroprotectant to treat TBI-induced brain injury via anti-apoptosis, and suppression of excessive activation of autophagy therefore has potential clinical therapeutic value for the treatment of TBI. http://dx.doi.org/10.1016/j.niox.2015.02.050
PP21 Interaction between three gaseous molecules in rat pancreatic acinar cells; with special reference to the participation in intracellular Ca2+ dynamics Amira Moustafa, Yoshiaki Habara Laboratory of Physiology, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan In pancreatic acinar cells, an increase in intracellular Ca2+ concentration ([Ca2+]i) is a key process for the induction of exocytosis. We have been attempting to find out intracellular machineries that are triggered by gaseous molecules, nitric oxide (NO), hydrogen sulfide (H2S) and carbon monoxide (CO) to induce or to modulate the increase in [Ca2+]i. In this presentation, by combining our recently obtained findings till now, an attempt was made to establish a putative overall cascade by which [Ca2+]i dynamics is regulated in rat pancreatic acinar cells. What we have demonstrated in our recent studies were: 1) All gaseous molecules induced the [Ca2+]i increase. 2) In the case of NO and H2S, such [Ca2+]i increase was found to be attributed to both Ca2+ entry and Ca2+ release. In contrast, it was indicated to be mainly via Ca2+ release in the case of CO. 3) The Gq–PLC–IP3–IP3R cascade was implicated to be the main pathway accelerated by all gaseous molecules. 4) Both H2S and CO most likely stimulate the PI3K–Akt/PKB pathway that may subsequently activate eNOS and cause NO production. 5) eNOS localization was detected in the nuclei of acinar, centroacinar and endothelial cells by immunohistochemical examination. 6) The produced NO would consequently trigger the cGMP-dependent PKG pathway via the activation of sGC. 7) The sGC–cGMP–PKG cascade was suggested to be involved in the H2S-induced [Ca2+]i release as well, though it was not likely to function in the case of CO, namely, CO induces the [Ca2+]i
MnSOD. Interestingly, hypoxia induced a transient increase of MnSOD activity and mRNA expression in 30 min, followed by a final loss of MnSOD function, while the conjugate pre-treatment could timedependently improve MnSOD activity, as well as its gene expression. We also found the protective effects were mediated by STAT3 phosphorylation, for STAT3 inhibition and knockdown both attenuated the protective effects of the conjugate. Furthermore, burst of mitochondrial ROS (mROS), dispersion of mitochondrial membrane potential (ΔΨm) and fragmentation of myofibril were also ameliorated with the conjugate treatment; again, the cell-protective effects of the conjugate were canceled by the STAT3 inhibitor 5,15-DPP. Innovation and conclusion: We present a novel hydrogen sulfide donor with excellent cardio-protective potential and better watersolubility, and find the cardio-protective mechanisms. The conjugate suppresses mitochondrial ROS through activation of the STAT3– MnSOD pathway, which is a novel way of mitochondrial ROS elimination. Keywords: Leonurine–SPRC conjugate; acute myocardial infarction; mitochondrial ROS; cardiac protection; STAT3–MnSOD pathway http://dx.doi.org/10.1016/j.niox.2015.02.048
PP19 Hydrogen sulfide reduces histopathology and improves functional outcome after traumatic brain injury in mice Mingyang Zhang a,b, Xiping Chen a, Luyang Tao a a Department of Forensic Sciences, Soochow University, Suzhou, China b Department of Forensic Sciences, Nantong University, Nantong, China Introduction: Hydrogen sulfide (H2S) is a lipid-soluble, endogenously produced gaseous messenger molecule collectively known as gasotransmitter. Over the last several decades, gasotransmitters have emerged as potent cytoprotective mediators in various models of tissue and cellular injury. Objects: To investigate changes of H2S after traumatic brain injury and its possible role, a mouse traumatic brain injury (TBI) model was established. Methods: Expression of Cystathionine-β-synthase (CBS) mRNA as a H2S-producing enzyme in mouse brain was determined by reverse transcriptase–polymerase chain reaction (RT-PCR). This study examines the neuroprotective effects of Hydrogen sulfide by lesion volume, motor performance and Morris water maze test after TBI in mice. Results: From the results of RT-PCR, it was found that the expression of CBS was down-regulated in mouse brain cortex and hippocampus after brain injury. Western blot analysis revealed that CBS was present in normal mouse brain cortex and hippocampus. Hydrogen sulfide in the cortex and hippocampus exhibited dynamic changes after brain injury, in parallel with CBS mRNA and protein expression. Moreover, pretreatment with the H2S donor (NaHS) could protect the neuron against the injury induced by TBI. Noticeably, the H2S donor NaHS could reduce TBI-induced injury assessed with lesion volume. Conclusion: These data suggested that H2S may have a therapeutic potential against neuron damage and improves functional outcome. http://dx.doi.org/10.1016/j.niox.2015.02.049
PP20 Hydrogen sulfide offers neuroprotection on traumatic brain injury in mice through modulation of apoptosis- and autophagyregulating molecules Mingyang Zhang a,b, Xiping Chen a, Luyang Tao a a Department of Forensic Sciences, Soochow University, Suzhou, China b Department of Forensic Sciences, Nantong University, Nantong, China
S21
Introduction: Traumatic brain injury (TBI) is a serious public health problem affecting millions of people in the world. Hydrogen sulfide (H2S), a novel gaseous mediator, has been recognized as an important neuromodulator and neuroprotective agent in the central nervous system. Objects: To investigate the possible role of H2S in traumatic brain injury (TBI), a mouse TBI model was established. The present study was undertaken to study the effects of exogenous H2S on traumatic brain injury (TBI) and the underlying mechanisms. Methods: The effects of exogenous H2S on TBI were examined by using measurement of brain edema, behavior assessment, propidium iodide (PI) staining, and Western blotting, respectively. To investigate whether H2S can ameliorate motor and cognitive dysfunction after TBI, motor test and Morris water maze were performed. Results: Compared to TBI groups, H2S pretreatment reduced brain edema, improved motor performance and ameliorated performance in the Morris water maze test after TBI. Compared to the TBI group, H2S pretreatment could decrease the number of PI-positive cells in injured cortex, dentate gyrus, CA1 and CA3 regions. Immunoblotting results showed that H2S pretreatment reversed TBIinduced cleavage of caspase-3 and decline of Bcl-2, suppressed LC3II, Beclin-1 and Vps34 activation and maintained p62 level in injured cortex and hippocampus post TBI. Conclusions: The present study demonstrates that systemic administration of H 2 S ameliorates brain edema and behavioral symptoms in TBI models. H2S may serve as a neuroprotectant to treat TBI-induced brain injury via anti-apoptosis, and suppression of excessive activation of autophagy therefore has potential clinical therapeutic value for the treatment of TBI. http://dx.doi.org/10.1016/j.niox.2015.02.050
PP21 Interaction between three gaseous molecules in rat pancreatic acinar cells; with special reference to the participation in intracellular Ca2+ dynamics Amira Moustafa, Yoshiaki Habara Laboratory of Physiology, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan In pancreatic acinar cells, an increase in intracellular Ca2+ concentration ([Ca2+]i) is a key process for the induction of exocytosis. We have been attempting to find out intracellular machineries that are triggered by gaseous molecules, nitric oxide (NO), hydrogen sulfide (H2S) and carbon monoxide (CO) to induce or to modulate the increase in [Ca2+]i. In this presentation, by combining our recently obtained findings till now, an attempt was made to establish a putative overall cascade by which [Ca2+]i dynamics is regulated in rat pancreatic acinar cells. What we have demonstrated in our recent studies were: 1) All gaseous molecules induced the [Ca2+]i increase. 2) In the case of NO and H2S, such [Ca2+]i increase was found to be attributed to both Ca2+ entry and Ca2+ release. In contrast, it was indicated to be mainly via Ca2+ release in the case of CO. 3) The Gq–PLC–IP3–IP3R cascade was implicated to be the main pathway accelerated by all gaseous molecules. 4) Both H2S and CO most likely stimulate the PI3K–Akt/PKB pathway that may subsequently activate eNOS and cause NO production. 5) eNOS localization was detected in the nuclei of acinar, centroacinar and endothelial cells by immunohistochemical examination. 6) The produced NO would consequently trigger the cGMP-dependent PKG pathway via the activation of sGC. 7) The sGC–cGMP–PKG cascade was suggested to be involved in the H2S-induced [Ca2+]i release as well, though it was not likely to function in the case of CO, namely, CO induces the [Ca2+]i